JPH0122246B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a drug that exhibits strong antibacterial activity against acid-fast bacteria, more specifically, the general formula () (In the formula, R ₁ and R ₂ independently represent a hydrogen atom or a lower alkyl group, and X represents a halogen atom.)
Pyrido [1,2,3-de] [1,
4] An anti-acid mycobacterial agent containing a benzoxazine derivative or a salt thereof. It is well known that acid-fast bacteria include Mycobacterium tuberculosis and atypical mycobacteria. Excellent drugs such as (EB), rifampicin (REP), and kanamycin (KM) have been developed. However, the occurrence of bacteria resistant to these existing tuberculosis drugs has reduced the intended effects of existing drugs, and conventional drugs alone are no longer necessarily sufficient as tuberculosis treatment drugs. Therefore, there is a desire to develop drugs that are effective against existing tuberculosis drug-resistant bacteria, and exploratory research is underway, and to date a certain number of drugs that are effective against specific drug-resistant bacteria have been discovered. However, there is no drug to date that is broadly effective against all bacteria resistant to existing tuberculosis drugs, and there is a strong desire to develop a drug that exhibits such effects. On the other hand, no drug has been found so far that clearly acts effectively against atypical acid-fast bacteria, which is one type of acid-fast bacteria, and there is a strong desire to develop a drug that has such an effect. In order to respond to the above-mentioned background, the present inventors
We conducted extensive research to find a drug that has excellent effects against acid-fast bacteria. In other words, in order to exhibit antibacterial activity against acid-fast bacteria, an optimal balance between lipophilicity and hydrophilicity is required in the bacterial cell membrane permeability of the drug, and the presence of basic groups and acidic groups in the aromatic ring. Considering that it would be desirable to have a compound with a group of Heading Completing the Invention. The compound of the general formula () according to the present invention is a new compound, and its production method is shown by the following reaction formula. (In the formula, R ₁ , R ₂ and X are the same as above.) That is, the compound () is dimethyl sulfoxide,
Compound () can be obtained by heating a compound of formula () in a polar solvent such as sulfolane, dimethylformamide, dimethylacetamide, or water at room temperature to 200°C, preferably 70 to 150°C for 1 to 48 hours. The compound of formula () thus obtained can be converted into a salt with an inorganic or organic acid such as hydrochloric acid, sulfuric acid or methanesulfonic acid, or a sodium salt or potassium salt of a carboxylic acid, by a conventional method. The fact that the thus prepared compound of formula () exhibits excellent mycobacterial activity against mycobacteria is demonstrated by the fact that 9-fluoro-3-methyl-10-(4-methyl-1) is a representative example of the compound of formula (). -Piperazinyl)-7-
Oxo-2,3-dihydro-7H-pyrido [1,
2,3-de〕[1,4]benzoxazine-6-
Using carboxylic acid (hereinafter referred to as DL-8280),
The minimum inhibitory concentration (MIC, μ
g/ml) was confirmed. As disclosed in the Examples, the drug of the present invention exhibits a wide range of remarkable effects against conventional Mycobacterium tuberculosis resistant bacteria and atypical acid-fast bacteria. When administering the drug of the present invention, it can be processed into various dosage forms such as capsules, tablets, powders, injections, etc. using known formulation techniques, and the dosage is determined according to the disease. Although it varies depending on the type and administration method, it is usually in the range of 0.2 to 2.0 g/day, preferably 0.2 g/day.
~1.0 g/day is possible. Furthermore, all the compounds of formula () according to the present invention have low toxicity; for example, the LD ₅₀ of DL-8280 is 380 mg/Kg (mouse IV).
Therefore, it can be safely used as a medicine. Reference examples and examples are described below. Example 1 Effect of the compound of the present invention on acid-fast bacteria
MIC (Part 1) Preparation of drug (DL-8280); Make a 150 mcg/ml solution with 50% propylene glycol aqueous solution and dilute with water. Medium: Ogawa medium and Kirchner medium: In Ogawa medium, drug is added before medium solidification. twenty five~
Kirchner liquid medium contains 1.56mcg/ml, and 3.12~
Contains 0.78mcg. Bacterial inoculation; 10 ^-3 mg/ml MIC data

[Table] Example 2 Low concentration of drug (25-0.39 mcg/ml) in Ogawa medium
MIC was measured in the same manner as in Example 1 except that .

【table】

[Table] Reference Example 1 5.0 g of 2,4-dichloro-3-fluoronitrobenzene and 5.8 g of powdered potassium fluoride are added to 5 ml of dimethyl sulfoxide and stirred at 140-155°C for 4.5 hours. The solvent was removed under reduced pressure and the residue was partitioned between water and chloroform. The chloroform layer was washed with water and dried, and then the chloroform was distilled off to obtain 3.8 g of 2,3,4-trifluoronitrobenzene as an oil. 20g of this and 150ml of dimethyl sulfoxide
Stir and add 10% potassium hydroxide aqueous solution dropwise at 18-20°C. The mixture was further stirred at room temperature for 2 hours, 1 portion of water was added, and the mixture was shaken with chloroform. The aqueous layer is acidified with hydrochloric acid and extracted with chloroform. The extract is washed with water, dried, and the chloroform is concentrated. The residue was purified by silica gel chromatography to obtain 5.8 g of 2,3-difluoro-6-nitrophenol as a yellow oil. 5.8 g of this product was added to 5.0 g of monochloroacetone, 8.0 g of potassium carbonate, 0.8 g of potassium iodide, and 100 ml of acetone, and refluxed for 4 hours. Insoluble materials are filtered off, the solvent is distilled off, and the residue is partitioned between chloroform and water. The chloroform layer is washed with water, dried, and then the solvent is distilled off and the residue is treated with n-hexane to obtain 5.0 g of 2-acetonyloxy-3,4-difluoronitrobenzene as pale yellow-white crystals. Dissolve 7.1 g of this product in 200 ml of ethanol, add 14 ml of Raney nickel, and perform atmospheric pressure catalytic reduction. After filtering off the catalyst and distilling off the solvent, the residue was decolorized through a layer of silica gel to yield 5.1 g of 7,8-difluoro-2,3-dihydro-3-methyl-4H-benzoxazine as a pale yellow oil. You can get it. A mixture of 4.8 g of this and 5.3 g of diethyl ethoxymethylenemalonate is heated at 140-145° C. for 1 hour. After the raw materials disappeared, a small amount of ethanol was distilled off under reduced pressure, and 35 g of ethyl polyphosphate was added to the resulting oil.
was added, stirred for 1 hour at a bath temperature of 140-145℃, poured into ice water after cooling, and the precipitate was dissolved in chloroform 600ml (200×
3) Extract. The chloroform layer was partitioned with a 5% aqueous potassium carbonate solution and then with water, and then dried with Glauber's salt to give 9,10-difluoro-3-methyl-7-oxo-
2,3-dihydro-7H-pyrido [1,2,3-
de] [1,4] 5.1 g of white powder of benzoxazine-6-carboxylic acid ethyl ester (melting point 261°C)
get. Add 4.0g of this to concentrated hydrochloric acid-acetic acid (1:4)50
ml and reflux in oil for 3 hours. After cooling, the precipitated crystals are collected by filtration, thoroughly washed with water, washed with a mixture of ethanol and ether (1:4), and dried under reduced pressure to obtain 3.7 g of carboxylic acid in the form of transparent plate-like crystals (melting point >300°C). 9,10-difluoro-3-methyl-7-oxo-2,3-dihydro-7H-pyrido [1,2,3
-de] [1,4] 1.0 g of benzoxazine-6-carboxylic acid and 2.85 g of N-methylpiperazine were added to 15 ml of dimethyl sulfoxide, and the mixture was heated at 100 to 110°C for 12 hours.
Stir for an hour. The reaction mixture was dried under reduced pressure, 40 ml of water was added to the residue, and the mixture was extracted with chloroform. After drying the extract, the residue was recrystallized from ethanol to give 9-fluoro-3-methyl-10-(4-methyl-1-piperazinyl) as colorless needle crystals with a melting point of 250-257°C (decomposition). )-7-oxo-2,3
-dihydro-7H-pyrido [1,2,3-de]
[1,4]Benzoxazine-6-carboxylic acid 550
Get mg. Elemental analysis value C ₁₈ H ₂₀ FN ₃ O ₄ Calculated value C 59.82, H 5.58, N 11.63 Analysis value C 59.62, H 5.59, N 11.65 Reference example 2 9,10-difluoro-7-oxo-2,3-dihydro -7H-pyrid [1,2,3-de] [1,
4] Dissolve 133 mg of benzoxazine-6-carboxylic acid and 200 mg of N-methylpiperazine in 2 ml of dimethyl sulfoxide, and stir at 90-110°C for 6 hours. After cooling, the precipitate is collected by filtration, washed with methanol and water, and then recrystallized from ethanol to give 9-fluoro-10-(4-methyl- 1-piperazinyl)-
53 mg of 7-oxo-2,3-dihydro-7H-pyrido[1,2,3-de][1,4]benzoxazine-6-carboxylic acid is obtained. Elemental analysis value C ₁₇ H ₁₈ FN ₃ O ₄ Calculated value C 58.78, H 5.22, N 12.10 Analysis value C 58.27, H 5.38, N 12.04 Reference example 3 9,10-difluoro-7-oxo-2,3-dihydro -7H-pyrid [1,2,3-de] [1,
4] Add 534 mg of benzoxazine-6-carboxylic acid and 688 mg of piperazine to 10 ml of dimethyl sulfoxide.
and stir at 90-100℃ for 5 hours. After cooling,
The precipitate was collected by filtration, washed with methanol, and then recrystallized from water to give 9-fluoro-7-oxo-10- as pale yellow needle crystals with a melting point of 258-268°C (decomposition).
(1-Piperazinyl)-2,3-dihydro-7H-
385 mg of pyrido[1,2,3-de][1,4]benzoxazine-6-carboxylic acid is obtained. Elemental analysis value C ₁₆ H ₁₆ FN ₃ O ₄ Calculated value C 57.65, H 4.84, N 12.61 Analysis value C 57.25, H 4.79, N 12.69 Reference example 4 9,10-difluoro-3-methyl-7-oxo-2 ,3-dihydro-7H-pyrido [1,2,3
-de] [1,4] 1.5 g of benzoxazine-6-carboxylic acid and 1.9 g of anhydrous piperazine are dissolved in 30 ml of dimethyl sulfoxide and reacted for 3 hours at a bath temperature of 100 to 110°C. The solvent was distilled off under reduced pressure, and the residue was washed with chloroform and then with ether, then stirred with 50 ml of 5% aqueous methanol at room temperature for 2 hours, and the precipitate was collected by filtration and washed with a small amount of the same solvent and then with methanol to obtain the residue. Recrystallizing the white powder from ethanol yields 9-fluoro-3-methyl-7-oxo-10-(1-piperazinyl)- with a melting point of 260°C (decomposition).
2,3-dihydro-7H-pyrido [1,2,3-
910 mg of pale yellow-white crystals of de][1,4]benzoxazine-6-carboxylic acid were obtained. Elemental analysis value C ₁₇ H ₁₈ FN ₃ O ₄・1/2H ₂ O Calculated value C 57.30, H 5.37, N 11.79 Analysis value C 57.01, H 5.42, N 11.75 Reference example 5 2,4-dichloro-3-fluoro 10.5 g (0.05 mol) of nitrobenzene in dimethyl sulfoxide 30
ml, add 8 ml of 40% aqueous sodium hydroxide solution, and stir at a bath temperature of 60-70°C for 20 hours. After the reaction,
Add 200 ml of water, extract and remove unreacted raw materials with ether, acidify the aqueous layer with acetic acid, and extract with ether. After drying the extract over Glauber's salt, the solvent was distilled off, and the residue was purified by column chromatography using 100 g of silica gel (developing solvent: chloroform).
Chloro-2-fluoro-6-nitrophenol
Obtain 3.4g. 3 g (15.7 mmol) of the above phenol compound, 3 ml of chloroacetone and 300 mg of powdered potassium iodide are refluxed in 50 ml of acetone for 6 hours with vigorous stirring. After cooling, insoluble matters were removed by filtration, the filtrate was concentrated, and purified by column chromatography using 20 g of silica gel (developing solvent: chloroform) to obtain 2.5 g of oily 2-acetonyloxy-4-chloro-3-fluoronitrobenzene. get. 2.3g of the above 2-acetonyloxy compound
(7.9 mmol) was dissolved in 30 ml of ethanol, and 2 g of Raney nickel was added for catalytic reduction. After the reaction, the catalyst was removed by filtration, the filtrate was concentrated, and the residue was dissolved in 20 g of silica gel.
Purified by column chromatography (developing solvent: chloroform) to obtain oily 7-chloro-8-fluoro-3-methyl-2,3-dihydro-4H-1,
1.2 g of 4-benzoxazine is obtained. 1.11g of the above benzoxazine compound
(5.5 mmol) and 1.4 g (6.2 mmol) of diethyl ethoxymethylenemalonate are heated and stirred at a bath temperature of 130 to 140°C for 2 hours. After confirming the disappearance of the raw material benzoxazine compound by thin layer chromatography, 5 g of ethyl polyphosphate was added to the reaction mixture, and the mixture was reacted again at a bath temperature of 140° C. for 1 hour. After cooling, add 20ml of water,
Extract the precipitate with 150 ml of chloroform.
After drying the extract over Glauber's salt, the solvent was distilled off, and the residue was purified by column chromatography using 20 g of silica gel (developing solvent: 5% methanol-chloroform).
9-chloro-10-(fluoro-
3-methyl-7-oxo-2,3-dihydro-
1.2 g of ethyl 7H-pyrido[1,2,3-de][1,4]benzoxazine-6-carboxylate is obtained. Elemental analysis value C ₁₅ H ₁₃ ClFNO ₄ Calculated value C 55.31, H 4.02, N 4.30 Analysis value C 55.19, H 3.97, N 4.41 600 mg (1.8 mmol) of the above ethyl carboxylate compound
was dissolved in 5 ml of concentrated hydrochloric acid-acetic acid (1:4, v/v) and heated at a bath temperature of 120°C for 6 hours. After cooling, add 20 ml of water to the reaction solution, collect the precipitated crystals by filtration, wash thoroughly with water, and add ethanol-ether (4:1, v/o).
v) Mixture, then wash with ether and dry, melting point
9-chloro-10- as a transparent plate-like crystal at 300℃ or higher
Fluoro-3-methyl-7-oxo-2,3-dihydro-7H-pyrido [1,2,3-de] [1,
4] Obtain 495 mg of benzoxazine-6-carboxylic acid. Elemental analysis value C ₁₃ H ₉ ClFNO ₄ Calculated value C 52.45, H 3.05, N 4.71 Analysis value C 55.20, H 3.13, N 4.74 Suspend 150 mg (0.5 mmol) of the above carboxylic acid in 3 ml of dimethyl sulfoxide, and add 1-methyl Add 150 mg of piperazine and react at a bath temperature of 120 to 130°C for 6 hours. After cooling, the solvent was distilled off under reduced pressure, and the residue was washed with ether to remove soluble materials, and then purified by column chromatography using 7 g of silica gel (developing solvent: 5% methanol-chloroform, 10% methanol-chloroform). The eluate containing the target compound was collected and the solvent was distilled off, and the residue was recrystallized from ethanol to give 9-chloro-3-methyl-10-(4- Methyl-1
65 mg of benzoxazine-6-carboxylic acid is obtained. Elemental analysis value C ₁₃ H ₂₀ ClN ₃ O ₄ Analysis value C 57.22, H 5.34, N 11.12 Calculated value C 57.20, H 5.11, N 11.23

Claims (1)

[Claims] 1. General formula (In the formula, R ₁ and R ₂ independently represent a hydrogen atom or a lower alkyl group, and X represents a halogen atom.)
An anti-acid and anti-mycobacterial agent containing the compound represented by or a salt thereof. 2 9-Fluoro-3-methyl-10-(4-methyl-1-piperazinyl)-7-oxo-2,3-
Dihydro-7H-pyrido[1,2,3-de][1,
4] The drug according to claim 1, which contains benzoxazine-6-carboxylic acid or a salt thereof.